Validated using a knockout cell line
Recombinant
RabMAb

Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)

Knockout Tested Rabbit recombinant monoclonal Niemann Pick C1 antibody [EPR5209]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human. Cited in 11 publication(s).

Overview

  • Product name
    Anti-Niemann Pick C1 antibody [EPR5209]
    See all Niemann Pick C1 primary antibodies
  • Description
    Rabbit monoclonal [EPR5209] to Niemann Pick C1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Niemann Pick C1 aa 1250-1350 (C terminal). The exact sequence is proprietary.
    Database link: O15118

  • Positive control
    • WB: Wild-type HAP1 whole cell lysate; HEK-293, HepG2, THP-1, PC-3, 3T3L1, L6 and HEK-293T cell lysates; Rat liver and brain lysates. IHC-P: Human kidney and liver tissues; Mouse liver tissue; Rat cerebellum tissue. ICC/IF: Neuro-2a cells; SH-SY5Y control and PADK treated cells. Flow Cyt: Neuro-2a cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Applications

Our Abpromise guarantee covers the use of ab134113 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/200.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/2000 - 1/10000. Detects a band of approximately 180 kDa (predicted molecular weight: 142 kDa).
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/70.

Target

  • Function
    Involved in the intracellular trafficking of cholesterol. May play a role in vesicular trafficking in glia, a process that may be crucial for maintaining the structural and functional integrity of nerve terminals.
  • Involvement in disease
    Defects in NPC1 are the cause of Niemann-Pick disease type C1 (NPDC1) [MIM:257220]. A lysosomal storage disorder that affects the viscera and the central nervous system. It is due to defective intracellular processing and transport of low-density lipoprotein derived cholesterol. It causes accumulation of cholesterol in lysosomes, with delayed induction of cholesterol homeostatic reactions. Niemann-Pick disease type C1 has a highly variable clinical phenotype. Clinical features include variable hepatosplenomegaly and severe progressive neurological dysfunction such as ataxia, dystonia and dementia. The age of onset can vary from infancy to late adulthood. An allelic variant of Niemann-Pick disease type C1 is found in people with Nova Scotia ancestry. Patients with the Nova Scotian clinical variant are less severely affected.
  • Sequence similarities
    Belongs to the patched family.
    Contains 1 SSD (sterol-sensing) domain.
  • Domain
    A cysteine-rich N-terminal domain and a C-terminal domain containing a di-leucine motif necessary for lysosomal targeting are critical for mobilization of cholesterol from lysosomes.
  • Post-translational
    modifications
    Glycosylated.
  • Cellular localization
    Late endosome membrane. Lysosome membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Niemann Pick C1 protein precursor antibody
    • Niemann Pick disease, type C1 antibody
    • Niemann-Pick C1 protein antibody
    • NPC antibody
    • NPC1 antibody
    • NPC1_HUMAN antibody
    see all

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling Niemann Pick C1 with ab134113 at 1/70 followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500(green). Confocal image showing cytoplasmic staining on Neuro-2a cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 and ab150120(AlexaFluor®594 Goat anti-Mouse secondary) at 1/500  (red).

    The negative controls are as follows:
    -ve control 1 – ab134113 at 1/70 followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500.

  • All lanes : Anti-Niemann Pick C1 antibody [EPR5209] (ab134113) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : NPC1 (Niemann Pick C1) knockout HAP1 whole cell lysate
    Lane 3 : HEK293 whole cell lysate
    Lane 4 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 142 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab134113 observed at 180 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab134113 was shown to specifically react with Niemann Pick C1 in wild-type HAP1 cells as signal was lost in NPC1 (Niemann Pick C1) knockout cells. Wild-type and NPC1 (Niemann Pick C1) knockout samples were subjected to SDS-PAGE. Ab134113 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Cathepsin B/L inhibition causes NPC disease-like cholesterol accumulation in SH-SY5Y cells.

    Confocal microscopy of SH-SY5Y control and PADK treated cells. Cholesterol (filipin staining, white) and NPC1 (ab134113; green).

  • ab134113 staining Niemann Pick C1 in Neuro-2a (mouse neuroblastoma cell line) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/200. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabeled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • All lanes : Anti-Niemann Pick C1 antibody [EPR5209] (ab134113) at 1/10000 dilution (purified)

    Lane 1 : HepG2 (human liver hepatocellular carcinoma cell line) cell lysate
    Lane 2 : THP-1 (human monocytic leukemia cell line) cell lysate
    Lane 3 : HEK-293 (human epithelial cell line from embryonic kidney) cell lysate
    Lane 4 : PC-3 (human prostate adenocarcinoma cell line) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 142 kDa
    Additional bands at: 180 kDa (possible glycosylated form)



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • All lanes : Anti-Niemann Pick C1 antibody [EPR5209] (ab134113) at 1/10000 dilution (purified)

    Lane 1 : 3T3-L1 cell lysate
    Lane 2 : L6 (rat skeletal muscle cell line) cell lysate
    Lane 3 : Rat tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 142 kDa
    Additional bands at: 180 kDa (possible glycosylated form)



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded human liver with purified ab134113 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin embedded mouse liver with purified ab134113 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin embedded rat cerebellum with purified ab134113 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • All lanes : Anti-Niemann Pick C1 antibody [EPR5209] (ab134113) at 1/1000 dilution (unpurified)

    Lane 1 : 3T3 L1 cell lysate
    Lane 2 : L6 (rat skeletal muscle cell line) cell lysate
    Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) cell lysate
    Lane 4 : THP-1 (human monocytic leukemia cell line) cell lysate
    Lane 5 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
    Lane 6 : PC-3 (human prostate adenocarcinoma cell line) cell lysate
    Lane 7 : Rat liver lysate
    Lane 8 : Rat brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

    Predicted band size: 142 kDa
    Observed band size: 180 kDa
    why is the actual band size different from the predicted?

  • Immunohistochemical analysis of paraffin embedded human kidney tissue labelling Niemann Pick C1 with unpurified ab134113 at 1/50.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:
  • Causevic M  et al. BACE1-cleavage of Sez6 and Sez6L is elevated in Niemann-Pick type C disease mouse brains. PLoS One 13:e0200344 (2018). WB ; Mouse . Read more (PubMed: 29979789) »
  • Hughes MP  et al. AAV9 intracerebroventricular gene therapy improves lifespan, locomotor function and pathology in a mouse model of Niemann-Pick type C1 disease. Hum Mol Genet 27:3079-3098 (2018). Read more (PubMed: 29878115) »
See all 11 Publications for this product

Customer reviews and Q&As

Answer

Thank you for your inquiry.

Unfortunately, I don't think that any of the https://www.abcam.com/index.html?t=158346&pt=1 we have in our catalog is suitable for the detection of the yeast Niemann-Pick type C-related protein 1 (http://www.uniprot.org/uniprot/Q12200).
I compared the immunogen sequence of each antibodywith the yeast protein and the maximum score is only 35%.

I would like to recommend checking the Biocompare website which has an excellent antibody search facility that includes many suppliers. The link is: http://www.biocompare.com

I hope this information is nevertheless helpful to you. Please do not hesitate to contact me if you have any further questions in this regard.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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